CN108169389A - The assay method of 6-gingerol content in processing aids ginger juice - Google Patents
The assay method of 6-gingerol content in processing aids ginger juice Download PDFInfo
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- CN108169389A CN108169389A CN201711434384.8A CN201711434384A CN108169389A CN 108169389 A CN108169389 A CN 108169389A CN 201711434384 A CN201711434384 A CN 201711434384A CN 108169389 A CN108169389 A CN 108169389A
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- gingerol
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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Abstract
The present invention provides a kind of assay method of 6 gingerol contents in processing aids ginger juice, is y=578.88x+91.297, r according to regression equation through high performance liquid chromatography detection through being ultrasonically treated obtained sample twice using methanol as extracting solution==0.9999 calculates 6 gingerol contents in ginger juice.Inventive samples processing method is simple, and 6 gingerols in sample solution, at least in interior stabilization for 24 hours, method favorable reproducibility, precision is high, suits large area to popularize.
Description
Technical field
The invention belongs to traditional Chinese medicine ingredients detection technique fields, and in particular to 6-gingerol content in a kind of processing aids ginger juice
Assay method.
Background technology
Ginger is the fresh rhizome of Zingiber herbaceos perennial ginger (Zingiber officinale Roscoe), rhizome
In irregular bulk, surface yellowish-brown or taupe brown, gas is fragrant, and characteristic taste is pungent.The traditional Chinese medical science thinks that CORTEX ZINGIBERIS RHIZOMAE is pungent cool, controls skin
Edema, the row severe edema due to hypofunction of the spleen；Ginger juice pungent-warm, pungent scattered stomach cold strength is strong, is chiefly used in vomitting；Rhizoma zingiberis pungent-warm, middle benefit gas instigate cold, Hui Yangtong
Arteries and veins, warm malaria strength is big, and baked ginger acrid flavour hardship does not walk table in walking, and temperature is lower burnt cold；Baked ginger charcoal is warm-natured, and relatively homoiothermy point is cold；It simmers
The bitter temperature of ginger, relatively warming intestines and stomach is cold.The pungent and scattered temperature of ginger, Energy benefit taste are apt to middle benefit gas stopping nausea and vomiting by lowering the adverse flow of QI, and dehumidify the ruffian that disappears, cough-relieving apophlegmatic, with
Stopping nausea and vomiting by lowering the adverse flow of QI is length.Research shows that ginger can saliva stimulating, gastric juice and digestive juice secretion, increase gastrointestinal peristalsis, promote food
It is intended to；Reducing temperature of heatstroke prevention is refreshed oneself, disinfecting detoxifying, swelling and pain relieving, carsickness-proof, treats migraine, eliminates drunk, the facial dark sore of mitigation, anti-
Dandruff is controlled, alleviates waist shoulder pain, foot foulness eliminating and other effects.The substances such as contained gingerol have very strong anti-oxidant in ginger
And effect of scavenging radical, the elderly, which often eats ginger, to remove " age spot ".
Drug ginger system is the common method of tcm clinical practice, and medicinal material of the successive dynasties through ginger has as many as 90 kinds；Medicinal material ginger system can
Be mitigated or eliminated medicinal material toxicity or side effect, can priming channel tropism, can be synergistic.But the most medicine materical crude slice in the whole nation at present
Factory, processing and use for ginger juice, particularly to the preparation technology parameter of ginger juice and quality but without stringent specification and mark
It is accurate.Processing aids, particularly Auxiliary Liquid Material as the admixture during process of preparing, directly affect the quality of medicine materical crude slice, should give
Give the attention of height.The chemical composition of processing aids ginger juice includes polysaccharide, Zingibain, flavonoids, gingerol class, volatile oil
Multiple components are waited, wherein gingerol constituents are as the active constituent generally acknowledged in ginger juice, and content is usually as ginger juice quality
One of judging quota.Since ingredient is sufficiently complex in ginger juice, chromatographic peak is compared with horn of plenty in chromatogram, and ginger contains a large amount of fibre
Dimension element, is unfavorable for the detection of sample, thus in practical applications ginger juice quality control there are it is many problem of.
Invention content
In order to solve the problems in the prior art, the purpose of the present invention is to provide 6-gingerol contents in a kind of ginger juice
Assay method.
Unless otherwise specified, number of the present invention is parts by weight, and the percentage is mass percent.
The object of the present invention is achieved like this：
The assay method of 6-gingerol content in ginger juice of the present invention, which is characterized in that using following steps：The fresh ginger system of taking
For standby ginger juice in conical flask, addition methanol is appropriate, is ultrasonically treated 20-40min, then leaves heart 8- at 1900 turn -2200
12min by supernatant liquid filtering, leaves heart 8-12min with 1500-1700, takes supernatant, obtain test solution；Precision weighs
6-gingerol reference substance is appropriate, and methanol is added to be configured to 1.226mgmL-lReference substance solution；Use Phenomenex Gemini
C18 chromatographic columns, acetonitrile:Methanol:Water is 43:5:52 be mobile phase, flow velocity：1.0mL/min；Column temperature：25 DEG C, measure wavelength:
280nm；Then the content of the 6-gingerol in ginger juice is calculated according to the linear relationship of 6-gingerol and ginger juice.
Preferably, inventive samples are prepared as：Ginger juice 10mL is taken, is put in conical flask, adds in methanol 20mL, precision claims
It is fixed, 30min is ultrasonically treated, weightlessness is supplied, shakes up, take 10mL, 2000 leave heart 10min, take supernatant liquid filtering, take subsequent filtrate,
16000 turns, 10min is centrifuged, takes supernatant to get test solution.
Further, ginger juice of the present invention by 100g fresh gingers adds 100mL water to squeeze the juice obtained.
The assay method of 6-gingerol content in ginger juice of the present invention, which is characterized in that using following steps：Take fresh ginger,
100mL water is added to squeeze the juice obtained ginger juice according to 100g fresh gingers, then take ginger juice 10mL, put in conical flask, add in methanol 20mL, essence
It is close weighed, 30min is ultrasonically treated, weightlessness is supplied, shakes up, take 10mL, 2000 leave heart 10min, take supernatant liquid filtering, take continuous filter
Liquid, centrifuges 10min, takes supernatant to get test solution by 16000 turns；It is appropriate that precision weighs 6-gingerol reference substance, adds first
Alcohol is configured to 1.226mgmL-lReference substance solution；Use Phenomenex Gemini C18 chromatographic columns, acetonitrile:Methanol:Water
It is 43:5:52 be mobile phase, flow velocity：1.0mL/min；Column temperature：25 DEG C, measure wavelength:280nm；Then ginger is calculated according to the following formula
The content of 6-gingerol in juice：
Y=578.88x+91.297；
Wherein：Y represents integrating peak areas value, and x represents sample size.
Advantageous effect：
The present invention provides a kind of assay method of 6-gingerol content in processing aids ginger juice, using methanol as extracting solution, warp
It is ultrasonically treated twice and sample is made, be y=578.88x+91.297, r according to regression equation through high performance liquid chromatography detection==
0.9999 calculates 6-gingerol content in ginger juice.Inventive samples processing method is simple, and the 6- ginger in sample solution obtained
Capsaicin is at least in interior stabilization for 24 hours, and method favorable reproducibility, precision is high, suits large area to popularize.
Description of the drawings
Fig. 1 is the canonical plotting of 6-gingerol.
Specific embodiment
The present invention is specifically described below by specific embodiment, it is pointed out here that following embodiment is served only for this hair
It is bright to be further described, it is impossible to be interpreted as limiting the scope of the invention, the person skilled in the art of this field can root
Some nonessential modifications and adaptations are made to the present invention according to foregoing invention content.All raw materials of the present invention and reagent are commercially available
Product.
Test material and instrument
Instrument：1200 high performance liquid chromatograph of Agilent, containing quaternary pump, DAD detectors, column oven, autosampler, color
Compose work station (AGILENT companies of the U.S.)；Ten a ten thousandth electronic balances of Shimadzu AUW220D；FA1204C a ten thousandth electronics day
It is flat；Multiplex vavuum pump of circulating water type：III types of SHZ-D (Yuhua Instrument Co., Ltd., Gongyi City)；UPH-I-10T is excellent general ultrapure
Hydrophone (Chengdu Ultra Pure Science ＆ Technology Co., Ltd)；RJ-TGL-16C types supercentrifuge (Wuxi Rui Jiang Analytical Instrument Co., Ltd).
Reagent：Acetonitrile：Chromatographically pure (Fisher, USA)；Methanol：Chromatographically pure (Sigma, USA)：High purity water：It makes by oneself in this laboratory；It is other
Reagent is that analysis is pure.Reference substance：6-gingerol (lot number：140914) it is provided by Chengdu Puffy moral Bioisystech Co., Ltd.
Ginger source such as the following table 1.
The experiment of table 1 ginger source
Embodiment 1
Method and result
The preparation of sample and reference substance solution
It is prepared by test solution
O.4mL precision draws ginger juice, put in 25mL measuring bottles, absolute ethyl alcohol is added to dissolve and is settled to scale, is shaken up, and filters,
To obtain the final product.
It is prepared by reference substance solution
Precision weighs 6-gingerol 5.20mg, until in 25mL measuring bottles, adds in absolute ethyl alcohol and dissolves and be settled to scale, shake
It is even, then the accurate 2mL that draws put in 10mL measuring bottles, and absolute ethyl alcohol is added to be settled to scale, shake up to get.
Measure the selected of wavelength：Respectively by 6-gingerol reference substance solution and ginger juice test solution in 200~400nm waves
Length is scanned, and as a result the two has absorption maximum at 281nm.
2 linear relationship of embodiment is investigated
It is appropriate that precision weighs 6-gingerol reference substance, and methanol is added to be configured to 1.226mgmL-lSolution, it is accurate respectively to inhale
1,5,10,15,18,20 μ L are taken, sample introduction is detected successively, and testing conditions are chromatographic column：Phenomenex Gemini C18
(4.6mm × 250mm, 5um)；Flow velocity：1.0mL/min；Column temperature：25℃；The selection of mobile phase:Acetonitrile:Methanol:Water (43:5:
52)；Measure wavelength:280nm.With reference substance sample size (mgmL-l) it is abscissa, integrating peak areas value is ordinate, is drawn
Standard curve simultaneously returns processing, the results showed that, 6-gingerol is in 1.226~24.52mgmL-lRange linear relationship is good, returns
It is y=578.88x+91.297, r to return equation==0.9999.See Fig. 1 canonical plottings.6-gingerol in 3 ginger juice of embodiment
Assay
Using 6-gingerol as index ingredient, 6-gingerol contains in different batches ginger juice prepared by measure technique for fixing
Amount.
The preparation of sample：
Fresh ginger is taken, 100mL water is added to squeeze the juice obtained ginger juice according to 100g fresh gingers, then takes ginger juice 10mL, put conical flask
In, methanol 20mL is added in, accurately weighed, supersound process 30min supplies weightlessness, shakes up, takes 10mL, and 2000 leave heart 10min,
Supernatant liquid filtering is taken, subsequent filtrate is taken, 16000 turns, centrifuges 10min, take supernatant, obtain sample.
Sample detection：Testing conditions are chromatographic column：Phenomenex Gemini C18 (4.6mm × 250mm, 5um)；Stream
Speed：1.0mL/min；Column temperature：25℃；The selection of mobile phase:Acetonitrile:Methanol:Water (43:5:52)；Measure wavelength:280nm.
As a result it calculates：It is y=578.88x+91.297, r according to regression equation==0.9999 calculates.
It is prepared by 4 test solution of embodiment
With reference to the detection method of embodiment 2-3, influence of the Extraction solvent to testing result is investigated.
The investigation of 2 Extraction solvent of table
Go out from the experimental results, with methyl alcohol process sample, under the same terms, peak area is maximum.
Embodiment 5
With reference to the detection method of embodiment 2-3, influence of the extraction time to testing result is investigated.
The investigation of the investigation of 3 extraction time of table
Tests prove that the preparation method of sample is：Fresh ginger is taken, 100mL water is added to squeeze the juice according to 100g fresh gingers obtained
Then ginger juice takes ginger juice 10mL, puts in conical flask, add in methanol 20mL, and accurately weighed, supersound process 30min supplies weightlessness,
It shakes up, takes 10mL, 2000 leave heart 10min, take supernatant liquid filtering, take subsequent filtrate, 16000 turns, centrifuge 10min, take supernatant,
Obtained sample, 6-gingerol content highest.
Embodiment 6
With reference to the detection method of embodiment 3, precision test is carried out.Same test solution is taken, sample introduction 6 times measures 6-
The peak area of gingerol, RSD 0.67%.The result shows that the precision of instrument is good.Testing result is shown in Table 4.
4 precision of table is investigated
Embodiment 7
With reference to the detection method of embodiment 3, stability test is carried out.Take same test solution, respectively at the 0th, 2,4,
8th, 16 it, is measured the peak area of 6-gingerol for 24 hours, the RSD of 6-gingerol is 1.08%.The result shows that in sample solution
6-gingerol is at least in interior stabilization for 24 hours.Testing result is shown in Table 5.
5 study on the stability of table
Embodiment 8
With reference to the detection method of embodiment 3, repetitive test is carried out.Take same batch of sample (No. 8), 6 parts of operation repetitive, according to
Secondary sample introduction measures the integrated peak areas of 6-gingerol, calculates content.Testing result is shown in Table 6, as a result shows repeated good.
6 repeatability of table is investigated
Embodiment 10
With reference to the detection method of embodiment 3, respectively different markets are purchased with 10 batches of gingers, 6- in prepared ginger juice
The peppery content of ginger is measured, and every batch of sample measures twice, the results are shown in Table 7.
7 assay of table
6-gingerol in sample solution produced by the present invention is at least in interior stabilization for 24 hours, method favorable reproducibility, precision
Height suits large area to popularize.
Claims (4)
1. the assay method of 6-gingerol content in a kind of ginger juice, which is characterized in that using following steps：Take prepared by fresh ginger
Ginger juice adds in methanol in conical flask, is ultrasonically treated 20-40min, then leaves heart 8-12min at 1900 turn -2200, will be upper
Clear liquid filters, and leaves heart 8-12min with 1500-1700, takes supernatant, obtain test solution；Precision weighs 6-gingerol pair
It is appropriate according to product, methanol is added to be configured to 1.226mgmL-lReference substance solution；Use Phenomenex Gemini C18 chromatographies
Column, acetonitrile:Methanol:Water is 43:5:52 be mobile phase, flow velocity：1.0mL/min；Column temperature：25 DEG C, measure wavelength:280nm；Then
The content of the 6-gingerol in ginger juice is calculated according to the linear relationship of 6-gingerol and ginger juice.
2. the assay method of 6-gingerol content in ginger juice as described in claim 1, it is characterised in that：The test solution
It is prepared as：Ginger juice 10mL is taken, is put in conical flask, adds in methanol 20mL, accurately weighed, supersound process 30min supplies weightlessness, shakes
It is even, 10mL is taken, 2000 leave heart 10min, take supernatant liquid filtering, take subsequent filtrate, 16000 turns, centrifuge 10min, take supernatant, i.e.,
.
3. the assay method of 6-gingerol content in ginger juice as described in claim 1, it is characterised in that：The ginger juice is fresh by 100g
It is obtained that ginger adds 100mL water to squeeze the juice.
4. the assay method of 6-gingerol content in a kind of ginger juice, which is characterized in that using following steps：Fresh ginger is taken, according to
100g fresh gingers add 100mL water to squeeze the juice obtained ginger juice, then take ginger juice 10mL, put in conical flask, add in methanol 20mL, and precision claims
It is fixed, 30min is ultrasonically treated, weightlessness is supplied, shakes up, take 10mL, 2000 leave heart 10min, take supernatant liquid filtering, take subsequent filtrate,
16000 turns, 10min is centrifuged, takes supernatant to get test solution；It is appropriate that precision weighs 6-gingerol reference substance, and methanol is added to match
1.226mgmL is made-lReference substance solution；Use Phenomenex Gemini C18 chromatographic columns, acetonitrile:Methanol:Water is
43:5:52 be mobile phase, flow velocity：1.0mL/min；Column temperature：25 DEG C, measure wavelength:280nm；Then ginger juice is calculated according to the following formula
The content of middle 6-gingerol：
Y=578.88x+91.297；
Wherein：Y represents integrating peak areas value, and x represents sample size.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109224044A (en) * | 2018-09-30 | 2019-01-18 | 康美药业股份有限公司 | A kind of concocting method and product of medicinal material baked ginger medicine materical crude slice |
| CN109406682A (en) * | 2018-12-21 | 2019-03-01 | 广东方制药有限公司 | The UPLC characteristic spectrum construction method and detection method of ginger medicinal material |
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| CN105486771A (en) * | 2015-12-23 | 2016-04-13 | 成都普思生物科技股份有限公司 | Fingerprint spectrum detecting method of common goldenrop particle compound preparation |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109224044A (en) * | 2018-09-30 | 2019-01-18 | 康美药业股份有限公司 | A kind of concocting method and product of medicinal material baked ginger medicine materical crude slice |
| CN109406682A (en) * | 2018-12-21 | 2019-03-01 | 广东方制药有限公司 | The UPLC characteristic spectrum construction method and detection method of ginger medicinal material |
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